Orally delivered dsRNA induces knockdown of target genes and mortality in the Asian long-horned beetle, Anoplophora glabripennis

Ramesh Kumar Dhandapani; Jian Jun Duan; Subba Reddy Palli

doi:10.1002/arch.21679

Orally delivered dsRNA induces knockdown of target genes and mortality in the Asian long-horned beetle, Anoplophora glabripennis

Ramesh Kumar Dhandapani, Jian Jun Duan, Subba Reddy Palli

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

Abstract

The Asian long-horned beetle (ALB) Anoplophora glabripennis is a serious invasive forest pest in several countries, including the United States. Methods available to manage or eradicate this pest are extremely limited, but RNA interference (RNAi) technology is a potentially effective method to control ALB. In this study, we used sucrose feeding bioassay for oral delivery of double-strand RNA (dsRNA) to ALB larvae. ³²P-labeled dsRNA orally delivered to ALB larvae using the sucrose droplet feeding method was processed to small interfering RNA. Feeding neonate larvae with dsRNA targeting genes coding for the inhibitor of apoptosis (IAP), vacuolar sorting protein SNF7 (SNF7), and snakeskin (SSK) induced knockdown of target genes and mortality. Feeding 2 µg of dsRNA per day for 3 days did not induce a significant decrease in the expression of target genes or mortality. However, feeding 5 or 10 µg of dsRNA per day for 3 days induced a significant decrease in the expression of target genes and 50–90% mortality. Interestingly, feeding 2.5 µg each of dsIAP plus dsSNF7, dsIAP plus dsSSK, or dsSNF7 plus dsSSK per day for 3 days induced a significant decrease in the expression of both target genes and approximately 80% mortality. Our findings demonstrate that orally delivered dsRNA induces target gene knockdown and mortality in ALB neonate larvae and RNAi technology may have the potential for effective ALB control.

Original language	English
Article number	e21679
Journal	Archives of Insect Biochemistry and Physiology
Volume	104
Issue number	4
DOIs	https://doi.org/10.1002/arch.21679
State	Published - Aug 1 2020

Bibliographical note

Funding Information:
We would like to thank Jonathan Schmude, Cherie Keenan, Linda Saundra, Daria Tatman, and Ellen Apacio (USDA ARS BIIRU), and Jeff Howell of the University of Kentucky for assistance with the rearing of Asian long-horned beetles. We also would like to thank Laura Ruberg and Rachel Brown for reviewing earlier versions of the manuscript, and Jinmo Koo for assisting with radioactivity work. This study was funded by the USDA Farm Bill Section 10007 (FY2018-AP18PPQS&T00C179), National Institutes of Health (GM070559-14 and 1R21AI131427-01) and by the National Institute of Food and Agriculture, USDA, HATCH under 2351177000.

Funding Information:
We would like to thank Jonathan Schmude, Cherie Keenan, Linda Saundra, Daria Tatman, and Ellen Apacio (USDA ARS BIIRU), and Jeff Howell of the University of Kentucky for assistance with the rearing of Asian long‐horned beetles. We also would like to thank Laura Ruberg and Rachel Brown for reviewing earlier versions of the manuscript, and Jinmo Koo for assisting with radioactivity work. This study was funded by the USDA Farm Bill Section 10007 (FY2018‐AP18PPQS&T00C179), National Institutes of Health (GM070559‐14 and 1R21AI131427‐01) and by the National Institute of Food and Agriculture, USDA, HATCH under 2351177000.

Publisher Copyright:
© 2020 Wiley Periodicals, Inc.

Keywords

ALB
dsIAP
dsSNF7
feeding RNAi
pest control

ASJC Scopus subject areas

Physiology
Biochemistry
Insect Science

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10.1002/arch.21679

Cite this

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title = "Orally delivered dsRNA induces knockdown of target genes and mortality in the Asian long-horned beetle, Anoplophora glabripennis",

abstract = "The Asian long-horned beetle (ALB) Anoplophora glabripennis is a serious invasive forest pest in several countries, including the United States. Methods available to manage or eradicate this pest are extremely limited, but RNA interference (RNAi) technology is a potentially effective method to control ALB. In this study, we used sucrose feeding bioassay for oral delivery of double-strand RNA (dsRNA) to ALB larvae. 32P-labeled dsRNA orally delivered to ALB larvae using the sucrose droplet feeding method was processed to small interfering RNA. Feeding neonate larvae with dsRNA targeting genes coding for the inhibitor of apoptosis (IAP), vacuolar sorting protein SNF7 (SNF7), and snakeskin (SSK) induced knockdown of target genes and mortality. Feeding 2 µg of dsRNA per day for 3 days did not induce a significant decrease in the expression of target genes or mortality. However, feeding 5 or 10 µg of dsRNA per day for 3 days induced a significant decrease in the expression of target genes and 50–90% mortality. Interestingly, feeding 2.5 µg each of dsIAP plus dsSNF7, dsIAP plus dsSSK, or dsSNF7 plus dsSSK per day for 3 days induced a significant decrease in the expression of both target genes and approximately 80% mortality. Our findings demonstrate that orally delivered dsRNA induces target gene knockdown and mortality in ALB neonate larvae and RNAi technology may have the potential for effective ALB control.",

keywords = "ALB, dsIAP, dsSNF7, feeding RNAi, pest control",

author = "Dhandapani, {Ramesh Kumar} and Duan, {Jian Jun} and Palli, {Subba Reddy}",

note = "Funding Information: We would like to thank Jonathan Schmude, Cherie Keenan, Linda Saundra, Daria Tatman, and Ellen Apacio (USDA ARS BIIRU), and Jeff Howell of the University of Kentucky for assistance with the rearing of Asian long-horned beetles. We also would like to thank Laura Ruberg and Rachel Brown for reviewing earlier versions of the manuscript, and Jinmo Koo for assisting with radioactivity work. This study was funded by the USDA Farm Bill Section 10007 (FY2018-AP18PPQS&T00C179), National Institutes of Health (GM070559-14 and 1R21AI131427-01) and by the National Institute of Food and Agriculture, USDA, HATCH under 2351177000. Funding Information: We would like to thank Jonathan Schmude, Cherie Keenan, Linda Saundra, Daria Tatman, and Ellen Apacio (USDA ARS BIIRU), and Jeff Howell of the University of Kentucky for assistance with the rearing of Asian long‐horned beetles. We also would like to thank Laura Ruberg and Rachel Brown for reviewing earlier versions of the manuscript, and Jinmo Koo for assisting with radioactivity work. This study was funded by the USDA Farm Bill Section 10007 (FY2018‐AP18PPQS&T00C179), National Institutes of Health (GM070559‐14 and 1R21AI131427‐01) and by the National Institute of Food and Agriculture, USDA, HATCH under 2351177000. Publisher Copyright: {\textcopyright} 2020 Wiley Periodicals, Inc.",

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N1 - Funding Information: We would like to thank Jonathan Schmude, Cherie Keenan, Linda Saundra, Daria Tatman, and Ellen Apacio (USDA ARS BIIRU), and Jeff Howell of the University of Kentucky for assistance with the rearing of Asian long-horned beetles. We also would like to thank Laura Ruberg and Rachel Brown for reviewing earlier versions of the manuscript, and Jinmo Koo for assisting with radioactivity work. This study was funded by the USDA Farm Bill Section 10007 (FY2018-AP18PPQS&T00C179), National Institutes of Health (GM070559-14 and 1R21AI131427-01) and by the National Institute of Food and Agriculture, USDA, HATCH under 2351177000. Funding Information: We would like to thank Jonathan Schmude, Cherie Keenan, Linda Saundra, Daria Tatman, and Ellen Apacio (USDA ARS BIIRU), and Jeff Howell of the University of Kentucky for assistance with the rearing of Asian long‐horned beetles. We also would like to thank Laura Ruberg and Rachel Brown for reviewing earlier versions of the manuscript, and Jinmo Koo for assisting with radioactivity work. This study was funded by the USDA Farm Bill Section 10007 (FY2018‐AP18PPQS&T00C179), National Institutes of Health (GM070559‐14 and 1R21AI131427‐01) and by the National Institute of Food and Agriculture, USDA, HATCH under 2351177000. Publisher Copyright: © 2020 Wiley Periodicals, Inc.

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N2 - The Asian long-horned beetle (ALB) Anoplophora glabripennis is a serious invasive forest pest in several countries, including the United States. Methods available to manage or eradicate this pest are extremely limited, but RNA interference (RNAi) technology is a potentially effective method to control ALB. In this study, we used sucrose feeding bioassay for oral delivery of double-strand RNA (dsRNA) to ALB larvae. 32P-labeled dsRNA orally delivered to ALB larvae using the sucrose droplet feeding method was processed to small interfering RNA. Feeding neonate larvae with dsRNA targeting genes coding for the inhibitor of apoptosis (IAP), vacuolar sorting protein SNF7 (SNF7), and snakeskin (SSK) induced knockdown of target genes and mortality. Feeding 2 µg of dsRNA per day for 3 days did not induce a significant decrease in the expression of target genes or mortality. However, feeding 5 or 10 µg of dsRNA per day for 3 days induced a significant decrease in the expression of target genes and 50–90% mortality. Interestingly, feeding 2.5 µg each of dsIAP plus dsSNF7, dsIAP plus dsSSK, or dsSNF7 plus dsSSK per day for 3 days induced a significant decrease in the expression of both target genes and approximately 80% mortality. Our findings demonstrate that orally delivered dsRNA induces target gene knockdown and mortality in ALB neonate larvae and RNAi technology may have the potential for effective ALB control.

AB - The Asian long-horned beetle (ALB) Anoplophora glabripennis is a serious invasive forest pest in several countries, including the United States. Methods available to manage or eradicate this pest are extremely limited, but RNA interference (RNAi) technology is a potentially effective method to control ALB. In this study, we used sucrose feeding bioassay for oral delivery of double-strand RNA (dsRNA) to ALB larvae. 32P-labeled dsRNA orally delivered to ALB larvae using the sucrose droplet feeding method was processed to small interfering RNA. Feeding neonate larvae with dsRNA targeting genes coding for the inhibitor of apoptosis (IAP), vacuolar sorting protein SNF7 (SNF7), and snakeskin (SSK) induced knockdown of target genes and mortality. Feeding 2 µg of dsRNA per day for 3 days did not induce a significant decrease in the expression of target genes or mortality. However, feeding 5 or 10 µg of dsRNA per day for 3 days induced a significant decrease in the expression of target genes and 50–90% mortality. Interestingly, feeding 2.5 µg each of dsIAP plus dsSNF7, dsIAP plus dsSSK, or dsSNF7 plus dsSSK per day for 3 days induced a significant decrease in the expression of both target genes and approximately 80% mortality. Our findings demonstrate that orally delivered dsRNA induces target gene knockdown and mortality in ALB neonate larvae and RNAi technology may have the potential for effective ALB control.

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KW - dsIAP

KW - dsSNF7

KW - feeding RNAi

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Orally delivered dsRNA induces knockdown of target genes and mortality in the Asian long-horned beetle, Anoplophora glabripennis

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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